Genetic variation in Zfy final intron region on Y-chromosome of Canis aureus, Canis lupus and Vulpes vulpes and sex-determination for samples of three canid species

Abstract

The proportion of females and males in the wild can be a result of past ecological impacts on mammalian populations, and it can offer valuable insights into the evolutionary history, demographics, and behavior of mammalian species. We aimed to determine sex of road-killed samples belonging to three canid species (Canis aureus, C. lupus and Vulpes vulpes from Türkiye, and V. vulpes from Iran), and to reveal whether there was genetic variation within and among three species by sequencing the Zfy final intron region on Y–chromosome. When sex-determination testing was performed on 109 canid samples using a specific primer pair for the Zfx and Zfy genes, two specific DNA bands were detected in males while one DNA band was detected in females. The ratio of males was relatively lower than females. This indicated that there were clearly differences between females and males in terms of the band patterns. We also sequenced partially the Zfy final intron region for V. vulpes, C. aureus and C. lupus; 539 bp, 546 bp and 562 bp, respectively. When analyzed together the partial Zfy sequences, there was no variation among the sequences of each canid species and no haplotype was shared among three species. No phylogeographic structure was observed for each species throughout the sampling areas due to the partial Zfy final intron region highly being conserved. This might be resulted from gene flow due to an extensive male-biased dispersal presumably have homogenized the genetic variation in the Y–chromosome throughout the distributional areas of the canid species. In phylogenetic tree, the Canis species were clustered together and the Vulpes species were grouped in basale position and this was compatible with previous studies.